BACKGROUND

Rheumatoid arthritis (RA) is a common inflammatory disease of the joints. Due to importance of inflammation and oxidative stress in the pathogenesis of RA, drugs that have anti-oxidant and anti-inflammatory properties such as N-acetyl cysteine(NAC), can be effective as adjunctive therapy in patients with RA.

OBJECTIVE

The aim of this study has been to evaluate the effects of oral NAC on inflammatory cytokines and oxidative stress in patients with RA.

METHODS

Beside standard treatment, the NAC group (23 patients) received 600 mg of NAC twice daily and the placebo group (19 patients) received identical placebo twice daily for 12 weeks. Serum levels of total oxidant status (TOS), total antioxidant capacity (TAC), nitric oxide (NO), total thiol groups (TTG), malondialdehyde (MDA), tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR) were measured at baseline and at the end of study.

RESULTS

After 12 weeks treatment, results showed that in the NAC group, the serum level of MDA, NO, IL-6, TNF-α, ESR and CRP was significantly lower than that baseline. Also, serum level of TAC and TTG, as antioxidant parameters, increased significantly. However, only NO, MDA and TTG showed a significant difference in the NAC group as compared to the placebo group at the end of study.

CONCLUSIONS

According to the results of this study, oral NAC can significantly reduce the several oxidative stress factors and inflammatory cytokines. These results need to be confirmed in larger studies with considering clinical outcomes of RA patients.

The posttranscriptional modification of messenger RNA (mRNA) and transfer RNA (tRNA) provides an additional layer of regulatory complexity during gene expression. Here, we show that a tRNA methyltransferase, TRMT10A, interacts with an mRNA demethylase FTO (ALKBH9), both in vitro and inside cells. TRMT10A installs N¹-methylguanosine (m¹G) in tRNA, and FTO performs demethylation on N⁶-methyladenosine (m⁶A) and N⁶,2'-O-dimethyladenosine (m⁶A_m) in mRNA. We show that TRMT10A ablation not only leads to decreased m¹G in tRNA but also significantly increases m⁶A levels in mRNA. Cross-linking and immunoprecipitation, followed by high-throughput sequencing results show that TRMT10A shares a significant overlap of associated mRNAs with FTO, and these mRNAs have accelerated decay rates potentially through the regulation by a specific m⁶A reader, YTHDF2. Furthermore, transcripts with increased m⁶A upon TRMT10A ablation contain an overrepresentation of m¹G9-containing tRNAs codons read by tRNA^Gln(TTG), tRNA^Arg(CCG), and tRNA^Thr(CGT) These findings collectively reveal the presence of coordinated mRNA and tRNA methylations and demonstrate a mechanism for regulating gene expression through the interactions between mRNA and tRNA modifying enzymes.

Background: Rare cases of immune checkpoint inhibitor (ICI)-associated celiac disease (ICI-CeD) have been reported, suggesting that disruption of tolerance mechanisms by ICIs can unmask celiac disease (CeD). This study aims to characterize the clinicopathological and immunophenotypic features of ICI-CeD in comparison to ICI-associated duodenitis (ICI-Duo) and usual CeD.

Methods: A medical and pathological records search between 2015 and 2019 identified eight cases of ICI-CeD, confirmed by tTG-IgA. Nine cases of ICI-Duo, 28 cases of moderate CeD, as well as 5 normal controls were used as comparison groups. Clinical information was collected from the electronic medical records. Immunohistochemistry for CD3, CD8, T-cell receptor gamma/delta (γδ), programmed death ligand 1 (PD-L1), and programmed death 1 (PD-1) were performed, with quantification of intraepithelial lymphocyte (IEL) subsets in three well-oriented villi. CD68, PD-L1, and PD-1 were assessed as a percentage of lamina propria surface area infiltrated by positive cells. Statistical significance was calculated by the Student's t-test and Fisher's exact test.

Results: The eight patients with ICI-CeD (F:M=1:3) and nine patients with ICI-Duo (F:M=5:4) presented similarly with diarrhea (13/17) and abdominal pain (11/17) after a median of 1.6 months on ICI therapy. In patients with ICI-CeD, tTG-IgA ranged from 104 to >300 IU/mL. Histological findings in ICI-CeD and ICI-Duo were similar and included expansion of the lamina propria, active neutrophilic duodenitis, variably increased IELs, and villous blunting. Immunohistochemistry showed that the average number of IELs per 100 enterocytes is comparable between ICI-CeD and ICI-Duo, with increased CD3⁺ CD8⁺ T cells compared with normal duodenum but decreased γδ T cells compared with CeD. Average PD-L1 percentage was 9% in ICI-CeD and 18% in ICI-Duo, in comparison to <1% in CeD and normal duodenum; average PD-1 percentage was very low to absent in all cases (<3%). On follow-up, five patients with ICI-CeD improved on a gluten-free diet (GFD) as the sole therapeutic intervention (with down-trending tTG-IgA) while the other three required immunosuppression. All patients who developed ICI-Duo received immunosuppression with variable improvement in symptoms.

Conclusions: ICI-CeD resembles ICI-Duo clinically and histologically but shares the serological features and response to gluten withdrawal with classic CeD. Immunophenotyping of IELs in ICI-CeD and ICI-Duo also shows similar CD3, CD8, γδ T cell subsets, and PD-L1 populations, all of which differed quantitatively from usual CeD. We conclude that ICI-CeD is biologically similar to ICI-Duo and is likely a variant of ICI-Duo, but treatment strategies differ, with ICI-CeD often improving with GFD alone, whereas ICI-Duo requires systemic immunosuppression.

Keywords: costimulatory and inhibitory T-cell receptors; immunotherapy; inflammation.

Glossina pallidipes salivary gland hypertrophy virus (GpSGHV; family Hytrosaviridae) can establish asymptomatic and symptomatic infection in its tsetse fly host. Here, we present a comprehensive annotation of the genome of an Ethiopian GpSGHV isolate (GpSGHV-Eth) compared with the reference Ugandan GpSGHV isolate (GpSGHV-Uga; GenBank accession number EF568108). GpSGHV-Eth has higher salivary gland hypertrophy syndrome prevalence than GpSGHV-Uga. We show that the GpSGHV-Eth genome has 190 291 nt, a low G+C content (27.9 %) and encodes 174 putative ORFs. Using proteogenomic and transcriptome mapping, 141 and 86 ORFs were mapped by transcripts and peptides, respectively. Furthermore, of the 174 ORFs, 132 had putative transcriptional signals [TATA-like box and poly(A) signals]. Sixty ORFs had both TATA-like box promoter and poly(A) signals, and mapped by both transcripts and peptides, implying that these ORFs encode functional proteins. Of the 60 ORFs, 10 ORFs are homologues to baculovirus and nudivirus core genes, including three per os infectivity factors and four RNA polymerase subunits (LEF4, 5, 8 and 9). Whereas GpSGHV-Eth and GpSGHV-Uga are 98.1 % similar at the nucleotide level, 37 ORFs in the GpSGHV-Eth genome had nucleotide insertions (n = 17) and deletions (n = 20) compared with their homologues in GpSGHV-Uga. Furthermore, compared with the GpSGHV-Uga genome, 11 and 24 GpSGHV ORFs were deleted and novel, respectively. Further, 13 GpSGHV-Eth ORFs were non-canonical; they had either CTG or TTG start codons instead of ATG. Taken together, these data suggest that GpSGHV-Eth and GpSGHV-Uga represent two different lineages of the same virus. Genetic differences combined with host and environmental factors possibly explain the differential GpSGHV pathogenesis observed in different G. pallidipes colonies.

Commercially available electrical muscle stimulators (EMS) provide functional electrical stimulation and are interfaced with reciprocating gait orthosis (RGO). The system which has been developed is described here as an EMS-RGO. Advantages of the system include: medically prescriptable subsystems available from manufacturers, and commercially recommended subsystems for applications such as gait training. The system itself employs four EMS units worn on a belt. It is controlled by remote switches and is interfaced to electrodes placed over the quadriceps, hamstring and gluteal muscle groups of each leg. Two EMS units (for quadriceps stimulation) function primarily for stand-up and sit-down. Two other EMS units (for stimulation of the hip extensors) function primarily for ambulation. Each EMS unit is powered by a nine volt alkaline transistor battery which provides about 36 stand-uphs and sit-down's and approximately 3.1 km of walking before replacement is necessary. The system has been evaluated on a T-5 level paraplegic individual who sustained a motor complete lesion (Frankel Class B) of the spinal cord over seven years ago. It is emphasized that successful EMS-RGO walking exercise must be preceded by a physical conditioning programme of active physical therapy. New battery technology (such as lithium batteries) may improve the useful lifespan of the system, and new electrode technology (such as TTGs) may improve patient acceptance of the system.

OBJECTIVE

Recent studies from several countries have shown that coeliac disease (CD) is increasingly being diagnosed in adults, as the availability of new, accurate serologic tests has made screening in the general population possible. No data exist regarding the prevalence of CD in Greece. The aim of this study was the implementation of a serologic screening procedure for CD in the adult general population of Thessaly, an area of central Greece, using a novel diagnostic algorithm.

METHODS

The study included 2230 participants (1226 women, 1004 men, median age 46 years, range 18-80 years), selected by systematic random sampling, from the adult general population of Thessaly. All the serum samples were tested for total immunoglobulin A (IgA)-serum levels, to exclude IgA deficiency. Samples with total IgA within the normal range were tested for IgA antibodies against native human-tissue transglutaminase (anti-tTG); samples that were anti-tTG positive were tested for IgA antiendomysial antibodies (EmA). Samples from participants with selective IgA deficiency were examined for IgG antigliadin antibodies. Participants who were EmA-positive or antigliadin antibody-positive were referred for intestinal biopsy and human leucocyte antigen (HLA) typing.

RESULTS

No participant with selective IgA deficiency was detected. Four individuals tested positive for EmA, all of whom were biopsy-proven coeliacs. Therefore, the CD prevalence in this general population sample is 1 : 558 or 1.8 per 1000 (SE 0.13). The four new patients with abnormal histology (two men, two women) were aged between 18 and 35 years. Two of them were considered to be asymptomatic and two presented with a subclinical course. All four had the heterodimer HLA-DQ2.

CONCLUSIONS

This first serological screening study for CD in Greece has demonstrated that CD prevalence in Thessaly is among the lowest reported in Europe.

The gene for N-carbamyl-L-amino acid amidohydrolase was cloned from Bacillus stearothermophilus strain NS1122A into E. coli. This gene started with a TTG triplet and was predicted to encode a peptide of 409 amino acids, with a calculated molecular weight of 44,248. The deduced amino acid sequence shared moderate homology with that of the corresponding enzyme of Pseudomonas sp. strain NS671.

The localizations of two transglutaminases [factor XIIIa and tissue transglutaminase (tTG)] and their mRNAs were examined in human brain tissues from neurologically normal and Alzheimer disease (AD) cases, using immunohistochemical and in situ hybridization methods. In all cases, meningeal macrophages and ependymal macrophage/microglia were positive for factor XIIIa. The mRNA encoding factor XIIIa was detected in macrophages and microglia. As reported previously, intense staining with the antibody to factor XIIIa of a subset of microglia was seen in the parietal cortex in AD brains. Few or no microglia were found associated with classical senile plaques. In contrast, many labeled microglia were associated with primitive plaques. Further-more, most of these cells were mainly seen in the subpial cortical layer but were very rare in the hippocampus. On the other hand, few factor-XIIIa-positive microglia were found in the parietal cortices from non-neurological cases, but moderate numbers were found in their hippocampal tissues. TG and its mRNA were localized in astrocytes in all the cases. In AD, a few neurofibrillary tangles were positive to tTG. These results suggest that the subsets of microglia which express factor XIIIa may play some roles in the early phase of AD pathology.

OBJECTIVE

Celiac disease is an important cause of chronic diarrhea, failure to thrive, and anemia in children. Mode of presentation of celiac disease has changed in last few years. Study was conducted to determine the mode of clinical presentation of a large group of patients with celiac disease and whether there has been a change in the presentation with the time.

METHODS

A prospective study was conducted on 134 children diagnosed to be having celiac disease in the Pediatric Gastroenterology, PGIMER, Chandigarh, from July 1st 2006 to December 31(st) 2007. Their detailed clinical profile was recorded on a pretested proforma and all patients underwent hemogram, liver function tests, IgA anti-tissue transglutaminase (anti tTG), and upper gastro-intestinal endoscopy.

RESULTS

Major symptoms at presentation were diarrhea (54.5%), failure to thrive (52.2%), abdominal distension (41%), anemia (40%), pain abdomen (19.4%), vomiting (15.7%) and constipation (2.2% of cases). 60.4% of patients had short stature. Anemia was microcytic hypochromic in 79.1% of patients, and dimorphic in 20.9%. Serum transaminases were raised in 38.8% of cases. The mean serum anti tTG level was 164.24U/ml (Range 0-749 U/ml) and levels correlated with the severity of small intestinal damage on biopsy. 15 patients were negative for the serology but 8 out of them had IgA deficiency and all had histopathology suggestive of celiac disease.

CONCLUSIONS

Classical presentation of celiac disease is less commonly encountered these days probably related to the more widespread use of serologic testing and early recognition of atypical manifestations of celiac disease.

Hyper-expression of a secretory exoglucanase, Exg, encoded by the cex gene of Cellulomonas fimi was previously shown to saturate the SecYEG pathway and result in dramatic cell death of recombinant Escherichia coli (Z.B. Fu, K.L. Ng, T.L. Lam, W.K.R. Wong, Cell death caused by hyper-expression of a secretory exoglucanase in Esherichia coli, Protein Expr. Purif. 42 (2005) 67-77). We propose here that the cell lysate ratio (Pre/Mat RQ) of the unprocessed precursor Exg protein (Pre-Exg) and its processed mature product (Mat-Exg) reflects the capacity of E. coli to secrete Exg. A Pre/Mat RQ of 20/80, designated the "Critical Value," was an important threshold measurement. A rise in the Pre/Mat RQ triggered a mass killing effect. The use of various secretion signal peptides did not improve the viability of cells expressing high levels of Pre-Exg under strong tac promoter control. However, use of the weaker vegG promoter in conjunction with a change in start codon of the spa leader sequence from ATG to TTG in a pM1vegGcexL plasmid construct resulted in a high level (0.9 U ml(-1)) of excreted Exg in shake-flask cultures. This was 50% higher than the best result obtained from plasmid construct lacUV5par8cex, using the lacUV5 promoter and the ompA leader sequence. Variations in the excreted Exg activities were attributable to differences in the Pre/Mat RQ values of the induced cultures harboring pM1vegGcexL and lacUV5par8cex. These values were 18/82 and 10/90, respectively. Employing fed-batch cultivation in two-liter fermentors, an induced JM101(pM1vegGcexL) culture yielded 4.5 U ml(-1) of excreted Exg, which was over six fold greater that previously reported. Our results illustrate the successful application of the Pre/Mat RQ ratio as a guide to the attainment of a maximum level of secreted/excreted Exg.

BACKGROUND

The prevalence of irritable bowel syndrome (IBS) in the community is 10%-20% and have symptom based diagnostic criteria. Many symptoms of celiac disease (CD) with 1% prevalence in some communities can mimic IBS. Sensitive and specific serologic tests of CD can detect asymptomatic cases. The purpose of this study was to compare the level of anti-tissue-transglutaminase (tTG) IgA in IBS patients and controls group.

METHODS

This case-control study was performed at a University hospital in which 107 patients with IBS who met the Rome II criteria for their diagnosis were compared with 126 healthy age and sex-matched controls. Both groups were investigated for CD by analysis of their serum tTG IgA antibody with human recombinant antigen. Titers were positive containing over 10u/ml and borderline if they were between 4 and 10 u/ml.

RESULTS

86 percent of IBS patients were female. The mean antibody level was 0.837 u/ml in IBS group and 0.933 u/ml in control group without any significant difference.

CONCLUSIONS

Results of this study may intensify disagreement on the situation of CD in IBS patients.

BACKGROUND

Serum anti-actin IgA antibodies (AAA) were identified in patients with celiac disease (CD), and a close correlation emerged between the presence of AAA and mucosa damage, but test for AAA found in celiacs have a wide range of sensitivity and specificity values.

OBJECTIVE

To compare 1) the sensitivity and specificity of untreated, calcium-chelated and heated sera from 102 celiacs, 52 sick patients and 103 healthy controls in the determination of AAA, and 2) the reliability of AAA with anti-transglutaminase antibodies (anti-tTG) in diagnosing celiac disease and in predicting intestinal damage. The intestinal derived AAA was isolated by using the phage-display library technique.

RESULTS

Treated sera was significantly more sensitive than untreated (p=0.0001), and showed a significant correlation between AAA and the three degrees (3a, 3b, 3c) of intestinal damage (p=0.01). Sensitivity and specificity values of anti-tTG assay were higher than the AAA assay, and anti-tTG serum-concentration was only significantly correlated with more severe (3b and 3c) intestinal damage degrees. AAA isolated by phage display showed similar results of serum AAA in immunofluorescence assay.

CONCLUSIONS

Notwithstanding correlation between AAA and celiac disease, AAA assay, also after treatments, has little to offer in screening for CD compared to the well-established anti-transglutaminase assay.

BACKGROUND

Screening for celiac disease is based on the sequential evaluation of serologic tests and intestinal biopsy; an optimal screening protocol is still under investigation. The screening policy of one of the main healthcare providers in Israel (Maccabi) consists of measuring total immunoglobulin A and tissue transglutaminase IgA antibodies and confirming positive results by endomysial antibodies. For IgA-deficient patients antigliadin IgG is measured.

OBJECTIVE

To evaluate the use of tTGA as a first-level screening test in patients suspected of having celiac disease

METHODS

The results of tTGA and EMA tests over a 3 month period were obtained from the laboratory computer. Letters were sent to the referring physicians of patients with positive tests, requesting clinical information and small intestinal biopsy results. tTGA was performed using an anti-guinea pig tTG-IgA enzyme-linked immunosorbent assay kit.

RESULTS

Overall, 2,505 tTGA tests were performed: 216 (8.6%) were tTGA-positive of which 162 (75%) were EMA-negative (group 1) and 54 (25%) EMA-positive (group 2.) Clinical information was obtained for 91 patients in group 1 and 32 in group 2. Small intestinal biopsy was performed in 33 (36%) and 27 patients (84%) in groups 1 and 2, respectively. Celiac disease was diagnosed in 4 biopsies (12%) in group 1 and 23 (85%) in group 2 (P < 0.0001). The positive predictive value was 45% for tTGA and 85% for EMA.

CONCLUSIONS

Symptomatic patients with positive tTGA and negative EMA have a low rate of celiac disease compared to those who are tTGA-positive and EMA-positive. Confirmation with EMA is advised when tTGA is performed as a first-level screening for suspected celiac disease.

BACKGROUND

Antiendomysial (EmA) and antitransglutaminase (anti-tTG) antibodies are the most specific indirect marker of coeliac disease (CD). It is not known whether the oral mucosa of patients with CD is able to produce these antibodies or not.

OBJECTIVE

To evaluate the ability of the oral mucosa of patients with CD to produce antibodies in an in vitro culture system.

METHODS

Twenty-eight patients with new diagnosis of CD (15 adults and 13 children) and 14 adult subjects with other diseases (controls) were studied. All underwent oral mucosa biopsy and subsequent EmA and anti-tTG assays on the mucosa culture medium.

RESULTS

Sensitivity and specificity of EmA and anti-tTG assayed in the oral mucosa culture medium for CD diagnosis were 54% and 100% and 57% and 100%, respectively. The CD clinical presentation, such as the presence of oral mucosa lesions, did not influence the results of the EmA and anti-tTG assays in the oral mucosa culture medium. There was an association between positivity of antibodies and greater severity of the oral mucosa lymphocyte infiltration.

CONCLUSIONS

This study demonstrates that the oral mucosa contributes to EmA and anti-tTG production in untreated patients with CD.

The 16,299 bp long mitochondrial genome (mitogenome) of a tessaratomid bug, Eusthenes cupreus (Westwood), is reported and analyzed. The mitogenome represents the first sequenced complete mitogenome of the heteropteran family Tessaratomidae. The mitogenome of E. cuopreus is a typical circular DNA molecule with a total AT content of 74.1%, and contains 13 protein-coding genes (PCGs), 22 transfer RNA (tRNA) genes, two ribosomal RNA (rRNA) genes, and a control region. The gene arrangement is identical with the most common type in insects. Most PCGs start with the typical ATN codon, except that the initiation codon for COI is TTG. All tRNAs possess the typical clover-leaf structure, except tRNASer(AGN), in which the dihydrouridine (DHU) arm forms a simple loop. Six domains with 45 helices and three domains with 27 helices are predicted in the secondary structures of rrnL and rrnS, respectively. The control region is located between rrnS and tRNA(Ile), including some short microsatellite repeat sequences. In addition, three different repetitive sequences are found in the control region and the tRNA(Ile)-tRNA(Gln)-tRNA(Met)-ND2 gene cluster. One of the unusual features of this mitogenome is the presence of one tRNA(Gln)-like sequence in the control region. This extra tRNA(Gln)-like sequence is 73 bp long, and the anticodon arm is identical to that of the regular tRNA(Gln).

The cya genes, coding for adenylate cyclase, from Escherichia coli and Erwinia chrysanthemi B374 are compared after determination of a 3632 bp long nucleotide sequence of the hemC-cya region of E. chrysanthemi, encompassing the whole cya gene. In spite of a large divergence between the two organisms, especially visible in non coding regions, the amino acid sequence of the proteins are very similar, except at the very distal carboxyl end. Codon usage is different in the two organisms, and E. chrysanthemi tends to restrict translation to codons ending in G or C. Conservation of the translation initiation start region (including the poor ribosome binding site GGCG, and the TTG start codon), suggests that a specific protein synthesis process controls adenylate cyclase expression. Finally a palindromic unit, of primary sequence differing from the E. coli counterpart, borders the gene in E. chrysanthemi.

OBJECTIVE

To assess reproducibility of core laboratory performance and impact on sample size calculations.

BACKGROUND

Little information exists about overall reproducibility of core laboratories in contradistinction to performance of individual technicians. Also, qualitative parameters are being adjudicated increasingly as either primary or secondary end-points. The comparative impact of using diverse indexes on sample sizes has not been previously reported.

METHODS

We compared initial and repeat assessments of five quantitative parameters [e.g., minimum lumen diameter (MLD), ejection fraction (EF), etc.] and six qualitative parameters [e.g., TIMI myocardial perfusion grade (TMPG) or thrombus grade (TTG), etc.], as performed by differing technicians and separated by a year or more. Sample sizes were calculated from these results. TMPG and TTG were also adjudicated by a second core laboratory.

RESULTS

MLD and EF were the most reproducible, yielding the smallest sample size calculations, whereas percent diameter stenosis and centerline wall motion require substantially larger trials. Of the qualitative parameters, all except TIMI flow grade gave reproducibility characteristics yielding sample sizes of many 100's of patients. Reproducibility of TMPG and TTG was only moderately good both within and between core laboratories, underscoring an intrinsic difficulty in assessing these.

CONCLUSIONS

Core laboratories can be shown to provide reproducibility performance that is comparable to performance commonly ascribed to individual technicians. The differences in reproducibility yield huge differences in sample size when comparing quantitative and qualitative parameters. TMPG and TTG are intrinsically difficult to assess and conclusions based on these parameters should arise only from very large trials.

BACKGROUND Due to the increased prevalence of celiac disease in chromosomal anomalies and other congenital anomalies, this study was conducted to evaluate the seroprevalence of celiac disease (CD) in patients with congenital heart defects (CHD). METHODS This case-control study was done on 1002 children in two groups of CHD patients (n=402) and controls (n=600). The serum tissue transglutamianse (TTG) levels were investigated. The two groups were compared in terms of TTG IgA levels and p<0.05 was considered as the significant level. RESULTS The means of serum TTG IgA levels in children with CHD and the control groups were 19.17±46.67 and 7.77±10.02 u/mL respectively (p=0.001). After ANOVA analysis a significant difference between two cyanotic and acyanotic subgroups of cases and control groups was observed (p=0.000). The follow up tukey test showed only non-significant difference between the cyanotic and acyanotic cases. The frequency of TTG IgA with the consideration of 20 u/mL as cut-off point showed a significant association with groups (X2=28.31 and p=0.000). CONCLUSION According to the results the serum TTG IgA levels were significantly higher in patients with CHD than normal children and screening for CD in children with CHD is recommended.

OBJECTIVE

Genome-wide association study recently identified the chromosome 3q22.3 as a novel locus associated with coronary artery disease (CAD). This study was designed to identify the critical haplotype blocks within this region in Han Chinese populations.

METHODS

We selected 1920 CAD patients and healthy participants from Han Chinese and genotyped 22 single nucleotide polymorphisms (SNPs) spanning 150 kilobases (kb) chromosomal region flanking rs9818870, a SNP associated with CAD at 3q22.3 in Caucasian.

RESULTS

Seven SNPs were found to be strongly associated with CAD in females and clustered in two haplotype blocks of ESYT3 gene. This was validated in two geographically isolated case-control populations. The two blocks were 14 and 25kb long, respectively. In a combined haplotype analysis, the odds ratios (95% confidence interval, permuted P value) were 0.70 (0.58-0.83, 2×10(-5)) and 1.44 (1.20-1.72, 5×10(-5)) for haplotypes TTG and CCA in block 1 as well as 0.73 (0.61-0.87, 3×10(-4)) and 1.35 (1.13-1.62, 0.0013) for haplotypes TCG and CTT in block 2, respectively. ESYT3 was expressed in human lymphocyte, vascular endothelial cell, and smooth muscle cell. The risk factors including gender, obesity, hypertension, diabetes, and hyperlipidemia exhibited strong effects on the genetic contribution to CAD.

CONCLUSIONS

We identified two haplotype blocks of ESYT3 gene in 3q22.3 region that likely harbor functional variants, which cooperate with other risk factors and play a role in the pathogenesis of coronary artery disease in females.

OBJECTIVE

A serological screening assay for celiac disease (CD), designed to simultaneously detect IgA and IgG anti-tissue transglutaminase (a-tTG) and IgA and IgG deamidated gliadin peptide antibodies (a-DGP), was recently developed. In this study, we establish the performance of this assay.

METHODS

We enrolled 41 CD patients and 18 CD patients on gluten-free diets. The diagnosis of CD was based on histological and serological criteria, including concomitant positive serology tests (a-tTG, IgA anti-endomysial antibodies). As control population, we enrolled 169 subjects: 145 disease controls and 24 blood donors. In all cases, serum samples were tested for: IgA a-tTG, IgG a-tTG, IgA a-DGP, IgG a-DGP, IgA anti-endomysial antibodies (EMA), IgA and IgG for a-tTG and a-DGP in a single assay.

RESULTS

The new test, QUANTA Lite (™) h-tTG/DGP Screen, detects all IgA and IgG antibodies against atTG and a-DGP present in a sample. In our study, the test showed 100% sensitivity and 91.12% specificity.

CONCLUSIONS

This study showed additional value of the new h-tTG/DGP Screen assay, which proved superior to more conventional assays and can be considered the best initial test for CD. Further studies are necessary to determine whether combination of h-tTG/DGP Screen with IgA a-tTG or IgA a-DGP can be used to obviate the need for duodenal biopsy in high- and low-risk populations.

OBJECTIVE

To determine the relation between undiagnosed celiac disease and dyspepsia in the community.

METHODS

Patients presenting to the gastroenterology outpatient clinic of Mersin University Hospital, aged between 18 and 70 years and with no malignancy, malabsorption, chronic diarrhea, inflammatory bowel disease, diabetes mellitus, heart failure or renal failure, were asked to complete a questionnaire for functional bowel disease (based on Rome II criteria for irritable bowel syndrome and dyspepsia). The patients diagnosed with dyspepsia based on Rome II criteria formed the dyspepsia group and those with gastrointestinal complaints other than dyspepsia and irritable bowel syndrome formed the control group. Serum tissue transglutaminase antibody (anti-tTG) was determined in all patients. The patients with anti-tTG levels of >20U/ml underwent endoscopic duodenal biopsy.

RESULTS

The study included a total of 137 patients, of whom 69 (50.4%) were assigned into the dyspepsia group and 68 (49.6%) into the control group. Of 137 patients, 24 (17.5%) had an anti-tTG level of>> or =20U/ml: 14 in the dyspepsia group (20.3%) and 10 in the control group (14.7%), with no significant difference between the groups (p=0.39). Of the 24 patients positive for anti-tTG, 15 (64.5%) underwent endoscopy, and of these 15 patients, 8 (53.3%) had endoscopic duodenal biopsy. Biopsy revealed that of the 4 patients in the dyspepsia group, 3 (75%) had Marsh type 0 histology (IEL<40, normal crypt) and 1 (25%) had Marsh type 3a histology. Of the 4 patients in the control group, 3 (75%) had Marsh type 0 histology and 1 (25%) had Marsh type 3a histology. Histopathological examinations showed celiac disease in 2 out of the 8 patients (25%) positive for anti-tTG who underwent biopsy. Intention to treat analyses revealed that 1 of 69 patients in the dyspepsia group (1.44%) and 1 of 68 patients in the control group (1.47%) had celiac disease.

CONCLUSIONS

Celiac disease in this patient population had a high prevalence. Further studies with larger sample sizes are needed to confirm the relation between dyspepsia and celiac disease.

The complete mitochondrial genome (mitogenome) of Dolycoris baccarum (Hemiptera: Pentatomidae) has been sequenced and annotated in this study. This mitogenome is 16,549 bp in length with an A+T content of 73.4%, and contains the typical 37 genes that are arranged in the same order as that of the putative ancestor of hexapods. All protein-coding genes (PCGs) start with ATN codons except for cox1 that uses TTG as the initial codon. Eleven PCGs stop with termination codon TAA, and cox1 and cox2 have single T as the incomplete stop codon. All the transfer RNA genes have the typical clover leaf secondary structure, except for trnS1 (AGN) that lacks the dihydrouridine arm as known in many other metazoa. The control region is located between rrnS and trnI, and is composed of 479 bp of non-repeat region and 1401 bp of repeat region. This is the third completely sequenced mitogenome from the family Pentatomidae of Hemiptera.

OBJECTIVE

The aim of this study was to determine the impact on intra-articular healing of muscle tissue retained on tendon grafts used for anterior cruciate ligament (ACL) reconstruction.

METHODS

In an animal study on 40 New Zealand rabbits, a semi-tendon/semi-muscle graft (SSG) and a total tendon graft (TTG) were individually harvested from the Achilles tendons in each animal. After transecting the ACLs in both knees of each rabbit, SSG and TTG were randomly used on bilateral sides of the knee for ACL reconstruction. After 2, 4, and 8 weeks, functional scoring, gross observations, and histological evaluations of the repaired knees were performed (each time point; n = 10). Biomechanical testing was conducted on remaining animals at 8 weeks (n = 10).

RESULTS

At 2, 4, and 8 weeks after surgery, there were no statistically significant differences in functional scores between the SSG group and TTG group (n.s.). As healing progressed, skeletal muscle on the SSG was gradually absorbed with a corresponding decrease in graft diameter, compared to TTG, at each time point (P < 0.001). However, healing and incorporation of the intra-articular graft in the SSG were more apparent than those in the TTG, based on histology. The vascularity and cellularity in the center of the sample were significantly greater in the SSG group than the TTG group at all the time points (P < 0.01). At 8 weeks, the SSG group's ultimate failure load, yield load, and elongation at failure were significantly less than for the TTG group (P < 0.01). There were no significant differences in stiffness between the two groups with biomechanical testing (n.s.).

CONCLUSIONS

Results of this study indicate that muscle left on tendon grafts promotes intra-articular healing and remodeling of the graft in a rabbit model. However, excessive amounts of retained skeletal muscle weaken tendon graft's strength for ACL reconstruction. Preserving small amounts of muscle on tendon grafts is feasible for improving the biological success of ACL reconstruction in humans.

This study was performed to investigate the effect of of transdermal testosterone gel (TTG) on controlled ovarian stimulation (COS) and IVF outcomes and ovarian morphology according to pretreatment duration in poor responders. A total of 120 women were recruited for this pilot study. They were randomized into control, 2 weeks, 3 weeks or 4 weeks TTG treatment groups. For three TTG treatment groups, 12.5 mg TTG was applied daily for 2 weeks, 3 weeks or 4 weeks in preceding period of study stimulation cycle. After 3 weeks of TTG pretreatment, significant increase of antral follicle count (AFC) and significant decreases of mean follicular diameter (MFD) and resistance index (RI) value of ovarian stromal artery were observed (p=0.026, p<0.001, p<0.01, respectively). The total dose of rhFSH administered for COS significantly decreased after 3 and 4 weeks TTG treatment both compared with control group (p<0.001, p<0.001). The numbers of oocytes retrieved and mature oocytes were significanty higher in 3 and 4 weeks TTG treatment groups than control group (p<0.001, p<0.001 in the number of oocytes retrieved; p<0.001, p<0.001 in the number of mature oocytes). The clinical pregnancy rate and live birth rate were increased only in 4 weeks TTG treatment group compared with control group (p=0.030 and p=0.042, respectively). These data demonstrated that TTG pretreatment for 3 to 4 weeks increases AFC and ovarian stromal blood flow, thereby potentially improving the ovarian response to COS and IVF outcome in poor responders undergoing IVF/ICSI.